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Wheat salt-tolerant gene TaOPR and application thereof

A wheat salt-tolerant gene and gene technology, applied in the direction of DNA/RNA fragments, the use of vectors to introduce foreign genetic material, recombinant DNA technology, etc., can solve the problems that the role of OPR genes has not been reported, and achieve the effect of improving salt tolerance

Inactive Publication Date: 2011-07-13
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, some genes that can significantly improve the salt tolerance of plants have been found, but the role of OPR genes in the process of plant salt tolerance has not been reported yet.

Method used

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  • Wheat salt-tolerant gene TaOPR and application thereof
  • Wheat salt-tolerant gene TaOPR and application thereof
  • Wheat salt-tolerant gene TaOPR and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1, TaOPR clone

[0031] 1.1 Extraction of wheat Total RNA

[0032] 1. Put the tissue material into a liquid nitrogen pre-cooled mortar, and fully grind it into powder in liquid nitrogen;

[0033] 2. After the liquid nitrogen has evaporated, transfer it to a 2ml centrifuge tube immediately, add about 1ml of Invitrogen’s TRIzol extract for every 100mg of material, after melting, repeatedly suck and blow with a sample gun, shake and mix the sample vigorously, and make the sample Fully lyse and place at room temperature for 5 minutes;

[0034] 3. Add 0.2ml of chloroform (chloroform), vigorously shake and mix for 15 seconds, and place at room temperature for 10 minutes;

[0035] Centrifuge at 12000 rpm for 15 minutes at 4.4°C;

[0036] 5. Carefully suck out the upper aqueous phase with a pipette, add it to a new 1.5ml centrifuge tube, add 500 μl of isopropanol (1:1 volume), mix well, and settle at -20°C for 30 minutes or overnight;

[0037] 6.4°C, centrifuge ...

Embodiment 2

[0093] Embodiment 2, prokaryotic expression analysis

[0094] 2.1 Construction of prokaryotic expression vector

[0095] The expression vector used was Pet32a, and the recipient strain was DE3. select Hin d III and Eco RI performed double digestion on pET32a and the pMD18-T vector containing the target gene respectively, recovered the large fragment of the vector and the small fragment of the target gene, and used T 4 After ligation with DNA ligase, the competent cells of E. coli DH10B were transformed. After the recombinant was identified, the prokaryotic expression vector with the target gene was obtained, and then the recipient strain BL21 (DE3) was transformed into competent cells for protein expression.

[0096] 1. Digestion of pET32a and pMD18-T

[0097] Extract the plasmids of pET32a and pMD18-T by alkaline lysis method, take 10ul each for Hin dIII and Eco RI double enzyme digestion. The plasmid extraction method is as above, and the plasmid double enzyme di...

Embodiment 3

[0160] Embodiment 3, construction of plant expression vector (35S promoter)

[0161] 3.1 Construction of 35S promoter plant expression vector

[0162] Using the plant expression vector pSTART, select Sac I and Bam HI performed double enzyme digestion on pSTART and the pMD18-T vector containing the target gene respectively, and recovered the large fragment of the vector and the small fragment of the target gene respectively. 4 After ligation with DNA ligase, the competent cells of Escherichia coli DH10B were transformed, and the plant expression vector with the target gene was obtained after the recombinant was identified.

[0163] (1) Plasmid pSTART empty vector and pMD18-T Sac I and Bam HI double enzyme digestion

[0164] Extract pSTART empty vector and pMD18-T plasmid by alkaline lysis method, take 10 μg of each for enzyme digestion, and the enzyme digestion system is as follows:

[0165] Bam HI 1μl

[0166] Sac I 1μl

[0167] pSTART vector / pMD18-T plasmid 1...

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Abstract

The invention discloses a wheat salt-tolerant gene TaOPR, a plant expression vector pSTART-TaOPR or pXQUbi-TaOPR containing the gene, and application of the gene TaOPR and the expression vector thereof in salt-tolerant plants, particularly arabidopsis thaliana or common wheat. Proved by experiments, the salt tolerance of the transgenic plants is obviously improved.

Description

technical field [0001] The invention belongs to the technical field of biological genetic engineering, and in particular relates to a salt-tolerant gene—a gene TaOPR and its application. Background technique [0002] Soil salinization seriously affects crop yields. Especially with the development of industry, soil salinization is becoming more and more serious, which has become a social problem of global concern. my country has a large population, and soil salinization is more serious, which has become an important factor restricting my country's economic and social development. Therefore, in addition to alleviating soil salinization, cultivating new varieties of salt-tolerant crops has become a very urgent task at present. [0003] It is a technology with broad application prospects to use transgenic improved plant technology to transfer new traits into high-biomass plants, so as to develop new high-efficiency transgenic plant varieties and use them for planting in salin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82
Inventor 夏光敏董蔚
Owner SHANDONG UNIV
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